The identification of T cell antigens expressed by human tumors has led to the clinical trials designed to boost the host immune response against their tumors (reviewed by 1-4). Cancer vaccine strategies include antigenic peptides, peptide loaded dendritic cells, recombinant adenoviruses, vaccinia viruses, and plasmid DNA. Despite evidence of vaccination, very few clinical responses have been reported. Furthermore, those patients which had a clinical response generally have no evidence of vaccination. We have recently described a TCR gene transfer a treatment strategy designed to provide a source of autologous tumor reactive T cell to any patient regardless of their immune status. The primary goal of this proposal is to determine TCR gene modified T cells can be safely administered and have anti-tumor efficacy using an animal model for human melanoma. We will also determine how TCR affinity, tumor antigen vaccines, and IL-2 impacts on the safety and efficacy of TCR gene modified T cells in vivo. In addition to safety and efficacy, we will use using sensitive PCR assays we developed to address relevant questions regarding the fate of adoptively transferred T cells in tumor bearing mice which are not easily addressed in cancer patients. The successful completion of the proposed studies will enable us to design better immunotherapy treatment strategies for patients with advanced cancer.